The Gly-952 residue of Saccharomyces cerevisiae DNA polymerase α is important in discriminating correct deoxyribonucleotides from incorrect ones

Siripan Limsirichaikul, Masanori Ogawa, Atsuko Niimi, Shigenori Iwai, Takashi Murate, Shonen Yoshida, Motoshi Suzuki

研究成果: Article査読

7 被引用数 (Scopus)

抄録

Gly-952 is a conserved residue in Saccharomyces cerevisiae DNA polymerase α (pol α) that is strictly required for catalytic activity and for genetic complementation of a pol α-deficient yeast strain. This study analyzes the role of Gly-952 by characterizing the biochemical properties of Gly-952 mutants. Analysis of the nucleotide incorporation specificity of pol α G952A showed that this mutant incorporates nucleotides with extraordinarily low fidelity. In a steady-state kinetic assay to measure nucleotide misincorporation, pol α G952A incorporated incorrect nucleotides more efficiently than correct nucleotides opposite template C, G, and T. The fidelity of the G952A mutant polymerase was highest at template A, where the ratio of incorporation of dCMP to dTMP was as high as 0.37. Correct nucleotide insertion was 500- to 3500-fold lower for G952A than for wild type pol α, with up to 22-fold increase in pyrimidine misincorporation. The Km for G952A pol α bound to mismatched termini T:T, T:C, C:A, and A:C was 71- to 460-fold lower than to a matched terminus. Furthermore, pol α G952A preferentially incorporated pyrimidine instead of dAMP opposite an abasic site, cis-syn cyclobutane di-thymine, or (6-4) di-thymine photoproduct. These data demonstrate that Gly-952 is a critical residue for catalytic efficiency and error prevention in S. cerevisiae pol α.

本文言語English
ページ(範囲)19079-19086
ページ数8
ジャーナルJournal of Biological Chemistry
278
21
DOI
出版ステータスPublished - 23-05-2003
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生化学
  • 分子生物学
  • 細胞生物学

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